FOR STUDIES WITH ANIMALS ON CAMPUS

Transcription

1 GUIDELINES FOR FACULTY, STAFF AND STUDENTS FOR STUDIES WITH ANIMALS ON CAMPUS UNIVERSITY OF MARYLAND, BALTIMORE COMPARATIVE MEDICINE PROGRAM THE UNIVERSITY OF MARYLAND SCHOOL OF MEDICINE MEDICAL SCHOOL TEACHING FACILITY - ROOM G SOUTH PINE STREET BALTIMORE, MARYLAND (410) (FAX) July 2005

2 TABLE OF CONTENTS Page Telephone Numbers Position Statement i ii - iii Introduction - Veterinary Resources and Comparative Medicine Program 1-2 Accreditation and Memberships 2 Technical and Consultation Services 2-3 Veterinary Medical Services 3-4 Clinical Medicine Pathology Sources of Research Animals 4 Animal Ordering and Purchasing Procedures 5-6 Charge Authorization and Billing 7 Per Diem Charges 7 Animal Housing 8-9 Location and Access to Facilities Amphibian and Fish Holding Areas Satellite Facilities Policies, Procedures and Guidelines Caging Policy Information Quarantine and Stabilization of Animals Transport Breeding of Animals Pets Pathogen Testing of Rodent Biologics Media Coverage Monoclonal Antibody Production Experiments Involving Biohazards Survival Surgical Procedures Personal Protective Equipment Program for Workers with Macaque Monkeys Prolonged Restraint

3 Guidelines for Aseptic Surgery on Rodents Standard Immunization Methodology/Practices Protection of the Animal Facility from Outbreaks of Adventitious Pathogens Laws and Regulations NIH Principles for Use of Animals Institutional Review of Animal Protocols Grant Submission and Review - Required Information Training for Investigators and Staff TABLE OF APPENDICES App Institutional Animal Care and Use Committee's (IACUC) 1 Phone Numbers Protocol Review Form - IACUC 2 Guide to Drugs Commonly Used in Laboratory Animals 3 Tranquilizers Analgesics Injectable Anesthetics Inhalation Anesthetics Anesthetics, Analgesic and Tranquilizers For Unusual Species: Amphibia and Fish Anticholinergic Preanesthetics Antimicrobials 4 Useful Web Links 5 Sample Veterinary Resources Forms 6 Animal Census Sheet Charge Authorization Form Technical Services Request Form Animal Request Form (for all animal acquisitions) Guide to Handling Laboratory Animals 7

4 Fluid and Drug Administration 8 Blood Collection 9 Commonly Used Sites for Blood Withdrawal 10 Videotape List 11 Guidelines for the Recognition and Alleviation 12 of Pain and Distress in Rodents and Rabbits Occupational Health Program 13 Endpoints in Animal Study Proposals 14

5 Comparative Medicine Program TELEPHONE NUMBERS Academic programs (including veterinary resident training) Research collaboration (e.g. animal model development) Veterinary Resources and related special services Louis DeTolla, VMD, PhD, ACLAM - Director, Comparative Medicine and Chief Veterinary Resources Chief, Veterinary Surgical Services Chief, Veterinary Medical Services and VAMC Attending Attending Veterinarian Attending Veterinarian Deputy Director, Veterinary Resources Facility Manager Veterinary Resources (VR) Administrative offices Technical assistance Clinical calls for veterinary services Facility Manager Diagnostic pathology Purchase of animals FAX The Veterinary Resources business and administrative offices, located on the ground floor of MSTF, Rm. G-100 at 10 South Pine Street, are staffed from 8:00 a.m. to 5:00 p.m. daily. Emergencies During normal business hours, a veterinarian may be contacted at For after-hours emergencies, the veterinarian can be contacted by pager as follows: 2) Dial on a touch-tone phone. 3) Wait for three beeps after the telephone rings. 4) Enter the telephone number the veterinarian should use to call you. In case of a dialing error, press the * three times and begin again. 5) When you have entered the telephone number, press the # key to transmit your message. Wait for a fast busy signal then hang up. Your call will be returned as soon as possible. i

6 UNIVERSITY OF MARYLAND, BALTIMORE POSITION STATEMENT ON THE USE OF ANIMALS IN RESEARCH The University of Maryland, Baltimore believes the responsible use of laboratory animals is essential for biomedical research into the prevention and treatment of human and animal disease. We affirm the moral obligation of our scientists to carry out this research on behalf of mankind and animals. Millions of Americans are alive today, and live healthier and more productive lives because our nation's health care professionals are able to employ safe and effective treatments including vaccines, surgical procedures and other valuable therapeutic methods developed with animal research. These same medical advances, improving the quality of life for mankind, are also being used by veterinarians to save our cherished pets and companion animals, enhance the health of farm animals, and preserve a future for wildlife and endangered species. The benefits of animal research to human and animal health is virtually unchallengeable and is fully substantiated by the scientific literature. UM,B supports this essential research for the benefit of current and future generations. While we continue to seek other means of testing new medicines and techniques, animals continue to be the best model for researchers attempting to cure human disease. For the most part, alternatives to animal use such as tissue and cell cultures are useful as supplements to research, but have not entirely replaced the necessity for live animal testing. Computer modeling is also a valuable adjunct to research, but cannot replace the prudent use of animals. The University of Maryland, Baltimore, however, does believe in the three R's of research animal use whenever possible, replacing, reducing, and refining. Replacing of animals with cell cultures, or vertebrates with invertebrates whenever possible; reducing the numbers used by responsible experimental design, and improved statistical inferences; and refining techniques to eliminate any possible pain or discomfort. Researchers at UM,B share the public's concern about the responsible use of animals in research. Peer committees and stringent federal guidelines (Public Health Service Policy and Animal Welfare Act) require scientists to explore other means of experimentation before considering animal testing. All research employing vertebrates must be reviewed and approved in advance by UM,B's Institutional Animal Care and Use Committees to ensure that animal use is necessary and that high standards of humane care are observed. In addition to ensuring the judicious use of animals, the University administration and researchers share the responsibility to safeguard the welfare of laboratory animals. UM,B's animal facilities have the highest accreditation possible and are managed by highly qualified veterinarians who specialize in laboratory animal care. UM,B's facilities meet the strict guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC), International and inspections by the United States Department of Agriculture. ii

7 UM,B defends the right of free speech. However, our responsibilities of providing and advancing medical care to society demand that we do not capitulate to tactics of intimidation and violence which undermine our democratic traditions and threaten the principle of free scientific inquiry. Therefore, UM,B cannot tolerate such acts on University property and will not allow such acts to influence University policy. To the extent necessary, we will prosecute or discipline those who break the law or UM,B regulations. It is essential that we continue to preserve and protect the right of our researchers to pursue knowledge for those who wait for better therapies and treatments for disease and disability, and for the good of all human and animal kind. iii

8 INTRODUCTION Veterinary Resources (VR) All veterinary services at The University of Maryland, Baltimore campus are provided by Veterinary Resources. Veterinary Resources maintains oversight for the acquisition, care and use of all research animals on campus. All animal facilities and the care and use programs on campus are accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International (AAALAC) and meet federal laws and guidelines for the humane and appropriate care and use of laboratory animals. Animals are housed at nine locations; each of these facilities is maintained by a staff of experienced laboratory animal technicians. The primary mission of Veterinary Resources is to provide humane and scientifically appropriate care of research animals at The University of Maryland, Baltimore. The facilities and program of animal care and use are maintained in compliance with the Animal Welfare Act of 1966 and all subsequent revisions (regulated by the USDA), and Public Health Service (PHS) guidelines. Veterinary Resources provides service as economically as possible by having a centralized animal care staff and by wholesale purchase of feed, bedding, caging and husbandry supplies. PHS Resource Improvement Grants submitted by Veterinary Resources have also provided monies for large capital improvements on campus. Veterinary Resources is the service division of Comparative Medicine. Comparative Medicine Comparative Medicine was established as an Academic Program in 1989 by The Executive Committee of The School of Medicine. Faculty members in Comparative Medicine hold joint appointments in other departments. Areas of faculty expertise are in the fields of clinical laboratory animal medicine, surgery, comparative pathology, microbiology, immunology, genetics, and infectious diseases. Comparative Medicine faculty members conduct independent and collaborative research in a variety of fields. They are available for consultation on animal research protocols, laboratory animal management, and development of grant applications proposing the use of animals. Members participate in the teaching of medical students as well as graduate and postdoctoral students. We also provide postdoctoral residency and graduate training programs for veterinarians in the fields of Laboratory Animal Medicine and Comparative Pathology. As members of Veterinary Resources, these individuals provide veterinary supervision for husbandry and health care related to facilities management, diagnosis, treatment and prevention of intercurrent disease in research animals. 1

9 Institutional Animal Care and Use Committee Each school has an Institutional Animal Care and Use Committee (IACUC) whose members are appointed by the Dean. The committees review all proposed animal research and animal care and use to ensure compliance with the Animal Welfare Act and PHS Guidelines. ACCREDITATION The University of Maryland, Baltimore has been accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC) since This allows principal investigators to meet the requirements of category 1 on PHS grant and contract applications, i.e., this accreditation should be recognized in your grant applications that require in vivo research. MEMBERSHIPS The Program of Comparative Medicine maintains membership in the American Association for Laboratory Animal Science (AALAS). This organization provides educational materials, meetings, and information for professional development of staff in the specific area of laboratory animal medicine. AALAS also publishes Comparative Medicine and Contemporary Topics in Laboratory Animal Science, important journals of current developments in husbandry, animal care and health-related research. Veterinary Resources also belongs to, seeks guidance from, or is associated with the National Association for Biomedical Research, the Foundation for Biomedical Research, the Scientists Center for Animal Welfare and Americans for Medical Progress. These organizations act as spokespersons for the biomedical research community and provide education in the use of animals in research and teaching. All veterinarians in our program are specialty trained in laboratory animal medicine. TECHNICAL AND CONSULTATION SERVICES FOR RESEARCH PROCEDURES The staff of Veterinary Resources will provide technical services such as blood withdrawal, administration of anesthetics, animal transportation, etc. to support investigators in their research activities. Services may also be provided for surgery, pathology and radiology. Certain services are provided on a fee-for-service basis. Inquiries and prior arrangements for these services can be arranged by contacting our office. Through pre-research consultations, budgeting for these services can be included in research grant applications. 2

10 Pre-Research Consultation The Program, through its faculty, has as one of its missions the provision of information and advice regarding: a) Special caging or experimental techniques. b) Selection of appropriate animal species to carry out specific animal techniques. c) Animal models of human diseases. d) Anatomical and physiological peculiarities of animals used in research. e) Techniques of anesthesia, analgesia, chemical restraint, and dosages. f) Techniques of blood and other sampling and drug or chemical administration. g) Pathological and clinical effects of intercurrent animal disease. h) Estimates of animal purchase prices and future per diem rates. We encourage such consultations prior to the preparation of grant and contract applications. Clinical Medicine VETERINARY MEDICAL SERVICES When an investigator, research staff member, student, technician or any other person associated with institutional animal use believes an animal is abnormal, sick, in discomfort, or otherwise requiring aid, a call should be placed to Veterinary Resources along with the completion of an Animal Health Problem Report available through VR. A staff veterinarian will respond and take appropriate action in consultation with the investigator. It is essential that clinical calls be initiated by the investigator, student, fellow or technician at the earliest sign of the abnormality. Veterinary Resources maintains complete animal diagnostic laboratories, two aseptic surgical suites, treatment and radiographic facilities. 3

11 Pathology Veterinary Resources performs diagnostic pathology for the purpose of identifying intercurrent disease in the animal population and to assist investigators in identifying protocol related problems which affect animal health and impact on successful research endeavors. Inquiries should be directed to Dr. DeTolla or the diagnostic laboratory. Animals should be submitted to the necropsy room on the ground floor of the MSTF Building as soon as possible after death or preferably for euthanasia. Carcasses must not be frozen (refrigerate instead). A complete description of the animal's history should be included. A preliminary diagnosis will be available following the gross examination. A final diagnosis will follow as soon as possible after histopathology, microbiology, and other diagnostic procedures. SOURCES OF RESEARCH ANIMALS Animals used at UM,B are purchased from commercial suppliers and dealers. There are several commercial vendors who provide rodents, rabbits, dogs, cats, farm animals and non-human primates. Approval of sources is based on the health status, genetic quality, costs, vendor reputation, site-visits and reliability. Approved Vendors - School of Medicine (Rodents and Rabbits):Tom Morris, Jackson Labs, Charles River, Taconic, Zivic Miller (SPF only), Covance, National Institute of Aging, and National Cancer Research Center. Investigators with specific requests for animals from sources not currently approved must contact Veterinary Resources. Veterinary Resources will contact the suppliers, conduct health status screens and make appropriate management and husbandry decisions to make every effort to accommodate the investigators needs. No animals may be brought into the School of Medicine without prior consultation and approval by Veterinary Resources. When dealing with commercial suppliers who have multiple production colonies, it is often best to try to obtain animals from the same colony to prevent differences in biological response. No animal may be housed in UM,B facilities unless the animal ordering procedures are followed. The introduction of animals without authorization by Veterinary Resources places the facility and the research of other investigators at great risk for loss of animals and data which could jeopardize the results of research endeavors, publication and funding. Non-human primates are obtained from commercial sources, universities, public or private primate centers. Veterinary Resources can help in locating sources for non-human primates. Anyone planning to purchase non-human primates must discuss source, history, and health status with Dr. DeTolla. Veterinary Resources will make all arrangements for shipping. 4

12 ANIMAL ORDERING AND PURCHASING PROCEDURES SCHOOL OF MEDICINE All requests for research animals to be delivered to the School of Medicine or School of Pharmacy must be placed through Veterinary Resources. This arrangement is necessary, in part, to assure that the proper housing is available and to assist investigators in obtaining healthy animals at reasonable costs from safe, dependable sources. All arrangements for the acquisition, transportation and receipt of animals are handled by Veterinary Resources (even if animals are no cost). Veterinary Resources will provide purchase and animal care cost estimates. All animal-based research requires prior approval by the relevant Institutional Animal Care and Use Committee (IACUC). To assure this, all investigators will provide a current protocol number to Veterinary Resources at the time an order is placed. Investigators must make sure that the protocol number used is correct for the procedure and species to be used. Animal Request Forms may be obtained from the administrative offices, Room G-100, MSTF. The procedure for ordering animals has been developed with the UM,B Division of Procurement and Supply to minimize delay and paperwork for investigators. Animal orders will be placed in the following manner: An Animal Request form will be completed by the investigator and forwarded to Veterinary Resources. Be sure to have complete information on type, source, and numbers of animals, and the number of the protocol approved by the Institutional Animal Care and Use Committee. Also, be sure to complete the required USDA information. A FAS account number and an authorized signature is required before an order can be placed. Orders may be faxed, hand delivered or sent through the campus mail. Telephone orders are NOT accepted. Purchase requests must be received by Veterinary Resources by noon on the Thursday prior to the week delivery is desired to ensure the vendor can supply the number and type of animals requested, and to ensure that Veterinary Resources has the proper housing available. An additional order fee will be charged for orders received after 12:00 noon if delivery is desired for the following week. Federal Law (Animal Welfare Act) requires the University to annually report the numbers of animals used in research according to the following categories: 5

13 Number of animals to be used involving no pain or distress Number of animals to be used with appropriate anesthetic, analgesic, or tranquilizer administered to avoid pain or distress Number of animals to be used involving pain or distress without the administration of anesthesia, analgesics, or tranquilizers (requires specific justification and approval) Please be accurate in the reporting of these numbers on the Animal Request Form. The animal suppliers will be telephoned by Veterinary Resources and arrangements will be made for delivery. Any unusual problems with orders will be transmitted back to the investigator. Investigators will be billed a nominal fee to offset the cost of long-distance phone calls and the facsimile machine for each order placed. The non-faxed Animal Request Form used by Veterinary Resources consists of four copies. Keep the pink copy for your records and return the other copies to Veterinary Resources. The animals are delivered to the facility designated on the request form. Housing for the animals is assigned after the animals have been checked for accuracy of species and type and examined to ensure they are in good condition. Animals arriving without a Veterinary Resources purchase order number will not be accepted or allowed to enter the facility. The majority of animals ordered by investigators are housed where the investigator requests and are received when requested. Unless you are contacted by Veterinary Resources, you may assume your animals have arrived and are being housed. The simplest way for an investigator to determine whether the animals have arrived is to visit the appropriate facility. Investigators are encouraged to use the comments section of the animal purchase request form to provide any pertinent information. This is especially helpful when ordering pregnant animals or litters, or for animals with special requirements prior to delivery. It is to an investigator's advantage to allow as much time as possible for vendors to supply an order. Veterinary Resources will make every effort to comply with the requests of investigators but cannot be responsible for "last minute" orders that vendors are unable to supply. CHARGE AUTHORIZATION AND BILLING SCHOOL OF MEDICINE 6

14 Investigators must provide a University FAS account number on a Charge Authorization form (see Appendix 6) that is signed by the investigator and the responsible administrator. This authorizes Veterinary Resources to transfer funds from the investigator's account to cover the cost of animal care and services. Investigators will be responsible for notifying Veterinary Resources of any changes in funding or expiration dates. One copy of the billing statement will be mailed to the individual whose name and address appears on the Charge Authorization form. Research protocols approved by the IACUC must be on file before an account can be opened. PER DIEM CHARGES Billing for per diem is based on daily animal census. The animal census is kept on a "door sheet" (sample in Appendix 6) in each housing area. These census sheets are collected on the last day of the month and the charges are calculated for each investigator's account according to the number of animals, number of days and the specified rate for each species. Veterinary Resources personnel will add incoming animals. Investigators and their staff should mark out all animals that have been used if they are not to be returned to the room. Animals born in the facility are added to the census at weaning. It is the investigator's responsibility to verify the census and perform weekly headcounts on each of their door sheets and subtract animals on the day they are used. Questions or concerns regarding census counts should be brought to the attention of Veterinary Resources as promptly as possible. Per diem charges are calculated in accordance with the NIH Cost Analysis and Rate Setting Manual for Animal Resource Facilities. The most significant per diem charges include labor, feed and bedding, equipment maintenance costs and husbandry supplies. Each cost category is allocated on a "per-species" basis at the end of the month and individual charges are assigned. Per diems paid by the investigator are only a part of total animal care costs, which are supplemented by the Dean and the Comparative Medicine Program. When possible, per diem charges are assessed by the Dean's office prior to the start of each fiscal year. The updated costs are sent to department chairpersons. Increases and/or decreases are based on the projected cost of care for the ensuing fiscal year. Investigators are encouraged to contact Veterinary Resources for current per diem rates. Investigators are also encouraged to contact Veterinary Resources prior to filing a grant application to obtain a "best estimate" of future per diem rates. 7

15 ANIMAL HOUSING Location The following areas are used for the housing of research animals. All species cannot be housed in every area. The housing areas are: School of Medicine Animal Facilities Medical School Teaching Facility (MSTF) Bressler Research Building (BRB) Howard Hall (HH) Maryland Psychiatric Research Center (MPRC) Health Sciences Facility (HSF) Dental School Animal Facility School of Pharmacy Animal Facility Baltimore Veterans Affairs Medical Center - Animal Facility Maryland Biotechnology Center The facility selected for housing animals will generally be the most convenient to the investigators' laboratories, and consistent with available space, PHS requirements regarding species separation, and with other needs of the investigator. School of Medicine Faculty All investigators who house animals in the School of Medicine must have a primary School of Medicine Faculty appointment. All School of Medicine faculty with laboratories in the School of Medicine must house their research animals in the School of Medicine animal facilities. Access to Facilities Access to the animal housing facilities for investigators and their staff can be granted only after a University access card I.D. badge has been obtained from the Campus Police. See your administrative staff or the Campus Police to obtain a badge. 8

16 Once an approved I.D. badge has been obtained, a memorandum on the investigator's department stationery should be sent/delivered to Veterinary Resources, Rm. G-100 of the MSTF building. The memorandum should contain the following information: 1. Name of the person requiring access 2. The facility/floor to which the person requires access 3. Social security number 4. Number from the back of the University I.D. badge The Campus Police can update the card access system within 24 hours of a request from Veterinary Resources. The simplest way to determine whether access has been granted is to try the card reader. Investigators are granted access only to one location (e.g., MSTF or BRB/HH) to help prevent the transmission of adventitious pathogens between facilities. Unauthorized persons (including visitors, friends, and children) are not permitted in the facilities without the approval of Veterinary Resources. Veterinary Resources veterinary staff and professional colleagues, such as site visitors accompanying the veterinary staff, have unrestricted access to all areas of the UM,B campus where animals are held. Amphibian and Fish Holding Areas We prefer to house all amphibians within our centralized facilities. Sites for housing of fish outside the centralized facilities must meet minimum standards. In general these areas must conform to the guidelines written by the Canadian Council on Animal Care as such guidelines are recognized by AAALAC. The guidelines include monitoring of water quality, appropriate husbandry and handling practices, and appropriate use of anesthetic and euthanasia methods for various species. Written standard operating procedures, prepared by the investigator, should be available in all fish holding areas. Records of water monitoring, treatments, and husbandry procedures should also be maintained. Guidelines and reference materials are available through Veterinary Resources. Satellite Facilities If animals will be housed for more than 24 hours outside of the animal facility, the area must be formally approved as a satellite area. This will require adequate justification and approval by the IACUC. 9

17 POLICIES, PROCEDURES AND GUIDELINES Caging Policy Due to the cost of specialized caging and an ever increasing animal census, investigators are urged to include a request for caging in all grant applications requiring the use of animals. Veterinary Resources should be contacted prior to grant application to discuss the appropriate type and quantity of caging that might be required. Grant Application Requests for Special Caging Previous caging policy at UM,B has been geared to provide conventional housing for laboratory animals used in research. Current budget arrangements are not necessarily able to meet the needs of investigators for special housing requirements such as microisolators, and other nonconventional caging or unusual quantities of particular caging. To meet these special requirements of individual investigators, a caging policy, effective May 1, 1991 requests that those investigators who have specific requirements include a request for funding for non-conventional lab animal caging in all grant applications to the NIH, NSF, DOD, ACS and other extramural agencies. The request for special caging equipment must be appropriate for the proposed research to ensure that sufficient animal housing is available for the maximum possible census at any one time during the life of the grant. Depending on the research timetable, the caging may be purchased in different years of the grant. Investigators should contact Veterinary Resources to discuss their potential specialized caging needs prior to the submission of a grant application (x6-3540, x6-3547). To insure that caging design is compatible and inter-changeable with the UM,B facility, special caging will be purchased from known vendors on behalf of investigators by Veterinary Resources. This will be done in consultation with investigators to ensure research requirements are met. Quarantine and Stabilization of Animals All newly received animals must be allowed a stabilization period of at least 48 hours prior to their use. This permits the animals to adapt to their surroundings resulting in a more stable physiological and behavioral state. Studies indicate that mice have altered immune functions and elevated corticosteroid levels for 48 hours following shipment. 10

18 Quarantine for Primates - ALL primates entering the facility must undergo a quarantine period of 10 weeks. Quarantine for Rodents - Rodents received from other than approved sources are required to be quarantined and tested prior to release to investigators. Quarantine period is 4 to 6 weeks. Animal Transport between Buildings Animals should not be transported from one research building to another. Exceptional circumstances require approval by Veterinary Resources. Under no circumstances should animals other than rodents and rabbits be transported in a public elevator (freight elevators should be used). All animals that are transported should be covered in a manner to obscure public view and prevent exposure of others to animal allergens. Breeding of Animals Pets Any proposed breeding requires specific approval by the IACUC. Research animals are not pets and should not be removed from the facilities. Concerns have to do with public perception of an animal removed from a research setting, the fact that these animals have generally been procured with grant monies, liability of the University, and our need to document the disposition of research animals in accordance with federal law. Conversely, personal pets should not be brought onto campus for treatment, or otherwise. Veterinary Resources does not operate a clinic or provide service for pet animals. In addition, certain pet animals (especially mammals) can harbor and spread infectious diseases to animals within the research facility. Only pet fish are exempted from this policy. Pathogen Testing of Rodent Biologics Rodent origin cell lines, tumor lines or any other biologics (e.g., body fluids) passaged through live rodents have the potential to transmit a wide range of rodent pathogens. One example is mouse hepatitis virus (MHV) which can be devastating even as a subclinical infection as it can confound experimental results, especially in immunological studies. Another example is lymphocytic choriomeningitis virus (LCM) which will interfere with experimental data and result in 11

19 rodent morbidity and mortality and is also zoonotic (i.e., may cause disease in personnel). All animal biologics (any tissues, serum, cells, tumor, or other animal passaged materials) must be adventitious pathogen free prior to introduction into any animal. Veterinary Resources will test these biologics at no cost to the investigator. This usually involves a serological evaluation (e.g. rodent cell line injected into a naive mouse which is subsequently assayed for antibodies to a panel of murine viral antigens). Media Coverage Policy for Research Utilizing Animals It is understood that publicity for research is important at the University of Maryland, Baltimore, School of Medicine and therefore should be supported and encouraged. Media coverage of animal-based research must be conducted in a socially sensitive fashion and meet all federal regulations and guidelines regarding the humane care and use of animals. It is important that media coverage be presented in a positive light and not prone to misrepresentation. Also, it should be determined if it is truly necessary to exhibit the animals or demonstrate their use or care. Any investigator who is contacted by the media to provide information on any animal-based research should contact the Office of Media Relations (X6-7820) without delay. If it is anticipated that animals will be exhibited to the media or if a portion of the media coverage includes specific use and/or care of animals, then the Office of Media Relations will contact the Chief of Veterinary Resources and the IACUC Chair for their recommendations. The media event should not proceed until these persons (or their designees) are satisfied that all has been done to ensure that media coverage will be conducted appropriately. A representative from Veterinary Resources and/or the IACUC may be present during the event and have authority to make any necessary changes to protect the investigator and University from misrepresentation. Monoclonal Antibody Production Guidelines Monoclonal antibodies should be produced in tissue culture. Ascites production should be done only if tissue culture methods cannot be used or are inadequate. IACUC requires justification for ascites production. Tissue culture support for the production of monoclonal antibodies is available through Veterinary Resources. This is provided as a fee-for-service activity and includes production of 12

20 monoclonal supernatants in standard or serum - free media, roller bottle culture, or hollow-fiber generation for larger quantities of antibodies. All hybridomas must be Mouse Antibody Production (MAP) tested for adventitious pathogens (e.g. murine viruses, mycoplasma) that could interfere with experiments or cause morbidity/mortality in individual rodents and/or place the animal facility at risk. All testing is provided by Veterinary Resources free of charge. Experiments Involving Biohazards No hazardous agents (infectious, oncogenic, radioactive or chemical) may be used in any animal facility without being cleared by the IACUC. The use of radioactive materials requires additional approval by Radiation Safety (x6-6281). The Director of EHS is a member of our SOM IACUC and reviews all protocols for hazards along with the full committee. The Institutional BioSafety Committee (IBC) also reviews for role of recombinant DNA and Select Agents. If special caging and care are required, Veterinary Resources must be contacted well in advance (prior to submitting an animal protocol is recommended). The veterinary faculty and facilities personnel of Veterinary Resources are available for consultation and advice on matters relevant to animal housing, care and use when biohazard agents are proposed. At the campus level, The Office of Environmental Health and Safety (x6-7055) and the UM,B Biohazard Committee (IBC) oversee all experimentation and use of biohazardous agents. See the EHS handbook for further information at If radioactive substances, infectious organisms, toxic chemicals, or chemical carcinogens are to be used in-vivo, the following points must be addressed in your protocol to the IACUC: The IACUC form also requires the implementation of an IN Vivo hazardous reference sheet. ABL level (1, 2, or 3) for infectious agents. Concentration Route of administration Duration of exposure Length of time animals will be kept following exposure Room location where agent is administered Location of animal housing post exposure Method of animal disposal We must protect the health of our employees and others. Those who attend to the care and housing of your animals may not be familiar with the nature of hazards being used. Therefore, we require the necessary information on the agent to best safeguard those who work with the animals. (See Occupational Health Program) 13

21 Survival Surgical Procedures The Institutional Animal Care and Use Committees have set minimum standards for animal operating rooms and laboratories in which surgery is performed. The standards are based on the NIH Guide for the Care and Use of Laboratory Animals. Adherence to these guidelines is necessary for compliance with the standards of accreditation by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC). The standards are meant to ensure that surgical procedures are performed in an appropriate environment using good surgical techniques. These standards apply to all mammalian species, including rabbits, but excluding rodents. Aseptic technique (e.g., surgical gloves, mask and sterile instruments) should be used for surgical procedures on rodents such as rats and mice. Rodent surgical areas may be a separate room or portion of a room. The area should be clean and orderly and should not be used for any other purpose during the time of the surgical activity. Animal housing areas may not be used for surgical procedures. For all rabbits and higher species, designated survival surgical areas are required and should be used only for that purpose. Use of the area for other purposes such as office space and equipment and supply storage, except for surgical and research equipment, is not acceptable. Non-survival surgical procedures may be performed in general purpose laboratories provided the rules outlined below are followed: a) There must be no eating, drinking or smoking in the laboratory during the surgical procedure. b) All equipment and surfaces within the room must be kept clean. c) The surgical area should be free of non-essential equipment and supplies. The use of a survival surgical area for non-survival surgery is satisfactory, provided the rules regarding survival surgery are followed, and the room is properly sanitized following the procedure. Any other laboratory outside of those designated for survival surgery CANNOT be used for survival surgical procedures in animals other than rodents. Approved surgical areas are in the Medical School Teaching Facility and the Bressler Research Building. Aseptic Technique Aseptic technique, including aseptic preparation of the skin, sterilization of instruments, and wearing sterile gloves and masks (gowns and caps when appropriate), is necessary when performing survival surgery on all animals. The IACUC provides guidelines on aseptic surgery. Operating 14

22 suites should not be used for eating, drinking or smoking. Personnel performing the surgery should have formal training in operative procedures and aseptic technique or have acceptable career experience approved by the IACUC. Anesthetics and Analgesics Information concerning types, dosages and routes of administration of anesthetics and analgesics is available from members of the Program. Also see Appendix 3. Anesthetic techniques and use of post-operative analgesia should be in accordance with current methods in the literature and approved by the IACUC. Inquiries may be directed to Veterinary Resources. Records must be maintained of the anesthetics used, amounts, dates, procedures and animal species. If volatile anesthetic gases are used, a gas evacuation and scavenging system is necessary. Analgesic usage - rule of thumb - if any particular procedure performed on animals would cause pain or discomfort if performed on a human then the animal should receive similar consideration in the form of analgesics. Guidelines for the Recognition and Alleviation of Pain and Distress in Rodents and Rabbits are included in Appendix 12. Post-Operative Care The investigator is primarily responsible for postoperative care and record keeping for animals until the animal is eating, drinking, and behaving normally. Immediate postoperative care should be provided in a dedicated recovery area with close observation and the animal should not be left until it can reach sternal recumbency. In the days following the procedure, the animal should be monitored and observed, as necessary, to insure the animal is recovering normally. Analgesics should be administered if there is any question of pain; incisions should be checked daily; bandages should be changed when wet, soiled, or coming off; supportive treatment should be provided when indicated (parenteral fluids, antibiotics); and sutures should be removed (7-14 days generally). For all species (except rodent), individual surgical records and post procedural records must be made and kept by the investigator. Forms for recording of surgical procedures and post operative care are available through Veterinary Resources. This record keeping is necessary to document our compliance with the USDA Animal Welfare Act and PHS (NIH) requirements for post procedural care of animals. Multiple Major Surgical Procedure Major surgery is defined as surgery which penetrates and exposes a body cavity or produces substantial impairment of physical or physiologic function. Multiple major survival surgery is described as more than one major survival surgical procedure performed on a single animal. 15

23 Multiple major survival surgical procedures may be performed on the same animal only if they are related components of a project and have been reviewed and approved by the IACUC. Multiple major survival surgery on animals for economic purposes alone is not acceptable according to the NIH Guide. Euthanasia Proper methods of euthanasia (see AVMA Panel on Euthanasia - ) should be used at the termination of the experiment. The methods used should be documented. A record must be kept of the amounts, date, and animal species when restricted drugs such as sodium pentobarbital are used for euthanasia. Personal Protective Equipment Program for Workers with Macaque Monkeys Training is required on the use of protective equipment for all who work with macaques or macaque tissues. This training is provided by the Veterinary Resources (VR) veterinarians. Macaque monkeys present a risk to employees from natural infection with Herpes B virus (other terms used Cercopithecine herpes virus 1, Herpes virus simiae, and B-virus). This latent infection in macaques is commonly fatal to humans that become infected. Humans have been infected from bites, scratches, splashes to the eye and from tissues of macaque monkeys. It is the investigator=s responsibility to make sure that each person under their employ receives this training and to notify Veterinary Resources of new employees who need training. Prolonged Restraint Policy Brief physical restraint of animals for examination, collection of samples, and a variety of other clinical and experimental manipulations can be accomplished manually or with devices such as restraint stocks or squeeze cages. It is important that such devices be suitable in size and design for the animal being held and operated properly to minimize stress and avoid injury to the animal. Prolonged restraint of any animal, including the chairing of non-human primates, should be avoided unless essential to research objectives. Less restrictive systems, such as the tether system or the pole and collar system, should be used when compatible with research objectives. The following are important guidelines for the use of restraint equipment: o o Animals to be placed in restraint equipment should be conditioned to such equipment prior to initiation of the research. The period of restraint should be the minimum required to accomplish the research objectives. Prolonged restraint for any reason must be approved by the IACUC committee. 16

24 o o o Restraint chairs or similar devices are not to be considered "normal" methods of housing, although they may be required for specific research objectives. Restraint chairs or similar devices must not be used simply as a convenience to investigators in handling or managing animals. When such devices are used, their use must be specifically approved by the IACUC committee. Attention must be paid to the possible development of lesions or illnesses associated with restraint, including contusions, decubital ulcers, dependent edema, and weight loss. If these or other problems occur, veterinary care must be provided to treat the animal, which if necessary must be temporarily or permanently removed from the restraint device. Guidelines for Aseptic Surgery on Rodents The following guidelines are consistent with our accrediting organization s (AAALAC International) interpretation of the Animal Welfare Act regulations and PHS guidelines which provide satisfactory aseptic conditions as indicated below. Investigators who feel that their vertebrate animal experiments require significant exceptions to these guidelines will need to justify these exceptions in their animal protocol. Otherwise, investigators will be expected to follow these guidelines. 1. Surgery should be conducted on a clean, uncluttered lab bench or table surface. The surface should be wiped with a disinfectant before and after use and/or covered with a clean drape. 2. Hair should be removed from the surgical site with clippers or a depilatory. The surgical site should be treated first with an antiseptic scrub (chlorhexidine or povidone iodine scrub) and rinsed with alcohol. 3. All instruments should be sterilized, but the method of choice may be determined by the surgical instruments or devices being used. Steam sterilization may be accomplished by autoclaving at 121 o C for 20 minutes. Gas sterilization with ethylene oxide is an alternative for items which will not withstand high temperature. Acceptable techniques for cold sterilization include soaking in 2% glutaraldehyde for ten hours, in 8% paraformaldehyde for 18 hours, or in 6% stabilized hydrogen peroxide for six hours. 4. The surgeon should wash his/her hands with an antiseptic surgical scrub preparation and then aseptically put on sterile gloves. If working alone, the surgeon should have the animal anesthetized and positioned and have the first layer of the double-wrapped instrument pack opened before putting on sterile gloves. 5. The surgeon should wear a surgical face mask. A surgical cap and gown are 17

25 recommended, but not required. 6. Multiple surgeries present special problems. After the first surgery, the sterilized instruments may be kept in a cold sterilant. Hot bead sterilizers are also acceptable to use between animals. Sterile gloves should be changed between surgeries if the surgeon touches nonsterile surfaces. 7. The abdominal or thoracic body wall should be closed with absorbable suture material. The skin should be closed with staples or with a nonabsorbable suture material in a simple interrupted pattern. Skin sutures or staples should be removed 7 to 10 days after surgery. 8. Rodents should be recovered from anesthesia in a warmed environment. Because of the rodent=s high surface area to volume ratio, they are particularly susceptible to heat loss (and overheating). Prevention of heat loss is therefore most desirable and may be accomplished by the placement of a dry cloth or pad under the animal during surgery. Warm water circulating heating pads are ideal for providing supplementary heat, if needed. Electrical heating pads should not be used as they can cause thermal burns and subsequent dermal necrosis. Heat lamps may be useful, but a thermometer should be placed near the animal to determine appropriate temperatures. Recovery on clean, dry bedding is sufficient for short procedures. 9. Antibiotics are not given routinely after surgery unless justified by the investigator. Antibiotics should not be used as a substitute for proper aseptic technique. 10. Analgesics should be used with all surgeries and whenever the potential for pain exists. Pain in rodents may be identified by observing the animal=s reluctance to move about, eat or drink and vocalization with handling. It is best not to wait for evidence of pain before giving analgesics. Buprenorphine mg/kg SQ is a safe and effective analgesic for rodents and should be given at the time of surgery and then every 8-12 hours. Lidocaine applied topically to the skin incision provides preemptive analgesia and may be sufficient for minor surgical procedures. Questions about aseptic or surgical technique may be addressed to Veterinary Resources staff by calling REFERENCES: a. Guide for the Care and Use of Laboratory Animals, U.S. Dept. of Health and Human Services, Public Health Service, National Institutes of Health, Revised b. Simmons, B.P., "CDC Guidelines for the Prevention and Control of Nosocomial Infections," American Journal of Infection Control, Vol. 11, 18

26 No. 13, June c. Wright, E.M., et al., "Animal Pain: Evaluation and Control," LAB ANIMAL, Vol. 14, No. 4, May/June d. Harkness, J.E., Wagner, J.E., The Biology and Medicine of Rabbits and Rodents, 4th edition, Lea and Febiger, 1995, Philadelphia. e. Block, S.S., Disinfection, Sterilization, and Preservation, 3rd edition, Lea and Febiger, 1983, Philadelphia. 19

27 Standard Immunization Methodology/Practices Veterinary Resources - School of Medicine University of Maryland, Baltimore Initial Injection Shave site area RABBIT (polyclonal antibody production) Antiseptic prep, alcohol or betadine Complete Freund's Adjuvant (CFA) + protein 1:1 (total 1 ml volume) 0.1 ml SQ or IM per site 0.05 ml ID per site maximum 10 sites, at least one inch separation of sites Foot pad injections are not acceptable for any species. Boost(s) Incomplete Freund's Adjuvant + protein -- same regimen as above. Blood sampling: maximum amount 1% of body weight (i.e., 3 kg rabbit - 30 ml) at minimum 2 week intervals. Methods - either marginal ear vein or central artery, by venipuncture or cut down. Droperidol (2.5 mg/kg) is recommended as a vasodilator + tranquilizer. Topical xylene is acceptable, but requires thorough washing afterward. Terminal Exsanguination requires complete anesthesia (ketamine and xylazine at 50 mg/kg and 5 mg/kg respectively IP, recommended) and may be performed through ear vein/artery or by cardiac puncture. Approved method of euthanasia should follow, such as pentobarbital, 100 mg/kg. MICE - monoclonal antibody production If Pristane priming is used, the recommended amount is 0.2 ml. intraperitoneal. 20